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Cloning, Molecular Characterization, and mRNA Expression of the Thermostable Family 3 β-Glucosidase from the Rare Fungus Stachybotrys microspora

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Abstract

The filamentous fungus Stachybotrys microspora possess a rich β-glucosidase system composed of five β-glucosidases. Three of them were already purified to homogeneity and characterized. In order to isolate the β-glucosidase genes from S. microspora and study their regulation, a PCR strategy using consensus primers was used as a first step. This approach enabled the isolation of three different fragments of family 3 β-glucosidase gene. A representative genomic library was constructed and probed with one amplified fragment gene belonging to family 3 of β-glucosidase. After two rounds of hybridization, seven clones were obtained and the analysis of DNA plasmids leads to the isolation of one clone (CF3) with the largest insert of 7 kb. The regulatory region shows multiple TC-rich elements characteristic of constitutive promoter, explaining the expression of this gene under glucose condition, as shown by zymogram and RT-PCR analysis. The tertiary structure of the deduced amino acid sequence of Smbgl3 was predicted and has shown three conserved domains: an (α/β)8 triose phosphate isomerase (TIM) barrel, (α/β)5 sandwich, and fibronectin type III domain involved in protein thermostability. Zymogram analysis highlighted such thermostable character of this novel β-glucosidase.

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Acknowledgments

Fatma Abdeljalil is thanked for reviewing English language. Lamia Jmal-Hammami and Mosbeh Dardouri are thanked for their technical help. We thank Professors Raja Mokdad-Gargouri and Hafedh Belghith for fruitful discussion of scientific interpretations. This work was supported by Grants from the Ministry of Higher Education and Scientific Research, Tunisia.

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  1. Laboratoire de Valorisation de la Biomasse et Production de Protéines chez les Eucaryotes, Centre de Biotechnologie de Sfax, University of Sfax, Route Sidi Mansour, BP 1177, 3018, Sfax, Tunisia

    Salma Abdeljalil, Héla Trigui-Lahiani, Houcine Lazzez & Ali Gargouri

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  1. Salma Abdeljalil
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  2. Héla Trigui-Lahiani
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  3. Houcine Lazzez
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  4. Ali Gargouri
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Correspondence to Ali Gargouri.

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Abdeljalil, S., Trigui-Lahiani, H., Lazzez, H. et al. Cloning, Molecular Characterization, and mRNA Expression of the Thermostable Family 3 β-Glucosidase from the Rare Fungus Stachybotrys microspora . Mol Biotechnol 54, 842–852 (2013). https://doi.org/10.1007/s12033-012-9633-5

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